A parallel permeability assay of peptides across artificial membranes and cell monolayers using a fluorogenic reaction

2019 ◽  
Vol 17 (11) ◽  
pp. 2887-2891 ◽  
Author(s):  
Jumpei Morimoto ◽  
Rei Amano ◽  
Takahiro Ono ◽  
Shinsuke Sando
Keyword(s):  
Membrane Permeability ◽  
Artificial Membranes ◽  
Cell Monolayers ◽  
Parallel Evaluation ◽  
Fluorogenic Reaction ◽  
Permeability Assay

An alkyne tag and a fluorogenic reaction enabled the rapid parallel evaluation of membrane permeability of peptides across artificial membranes and cell monolayers.

scholarly journals Omp35, a New Enterobacter aerogenes Porin Involved in Selective Susceptibility to Cephalosporins

2004 ◽  
Vol 48 (6) ◽  
pp. 2153-2158 ◽  
Author(s):  
Charléric Bornet ◽  
Nathalie Saint ◽  
Lilia Fetnaci ◽  
Myrielle Dupont ◽  
Anne Davin-Régli ◽  
...  
Keyword(s):  
Membrane Permeability ◽  
Outer Membrane ◽  
Antibiotic Susceptibility ◽  
Enterobacter Aerogenes ◽  
Artificial Membranes ◽  
Specific Location ◽  
Outer Membrane Permeability ◽  
Constriction Region ◽  
High Conductance

ABSTRACT In Enterobacter aerogenes, β-lactam resistance often involves a decrease in outer membrane permeability induced by modifications of porin synthesis. In ATCC 15038 strain, we observed a different pattern of porin production associated with a variable antibiotic susceptibility. We purified Omp35, which is expressed under conditions of low osmolality and analyzed its pore-forming properties in artificial membranes. This porin was found to be an OmpF-like protein with high conductance values. It showed a noticeably higher conductance compared to Omp36 and a specific location of WNYT residues in the L3 loop. The importance of the constriction region in the porin function suggests that this organization is involved in the level of susceptibility to negative large cephalosporins such as ceftriaxone by bacteria producing the Omp35 porin subfamily.

scholarly journals Biomimetic Artificial Membrane Permeability Assay over Franz Cell Apparatus Using BCS Model Drugs

Pharmaceutics ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 988
Author(s):  
Leonardo de Souza Teixeira ◽  
Tatiana Vila Chagas ◽  
Antonio Alonso ◽  
Isabel Gonzalez-Alvarez ◽  
Marival Bermejo ◽  
...  
Keyword(s):  
Membrane Permeability ◽  
Electron Paramagnetic Resonance Spectroscopy ◽  
Oral Drug ◽  
Resonance Spectroscopy ◽  
Artificial Membrane ◽  
Apparent Permeability ◽  
Franz Cell ◽  
Bcs Model ◽  
Fda Guidance ◽  
Permeability Assay

A major parameter controlling the extent and rate of oral drug absorption is permeability through the lipid bilayer of intestinal epithelial cells. Here, a biomimetic artificial membrane permeability assay (Franz–PAMPA Pampa) was validated using a Franz cells apparatus. Both high and low permeability drugs (metoprolol and mannitol, respectively) were used as external standards. Biomimetic properties of Franz–PAMPA were also characterized by electron paramagnetic resonance spectroscopy (EPR). Moreover, the permeation profile for eight Biopharmaceutic Classification System (BCS) model drugs cited in the FDA guidance and another six drugs (acyclovir, cimetidine, diclofenac, ibuprofen, piroxicam, and trimethoprim) were measured across Franz–PAMPA. Apparent permeability (Papp) Franz–PAMPA values were correlated with fraction of dose absorbed in humans (Fa%) from the literature. Papp in Caco-2 cells and Corti artificial membrane were likewise compared to Fa% to assess Franz–PAMPA performance. Mannitol and metoprolol Papp values across Franz–PAMPA were lower (3.20 × 10−7 and 1.61 × 10−5 cm/s, respectively) than those obtained across non-impregnated membrane (2.27 × 10−5 and 2.55 × 10−5 cm/s, respectively), confirming lipidic barrier resistivity. Performance of the Franz cell permeation apparatus using an artificial membrane showed acceptable log-linear correlation (R2 = 0.664) with Fa%, as seen for Papp in Caco-2 cells (R2 = 0.805). Data support the validation of the Franz–PAMPA method for use during the drug discovery process.

scholarly journals Paraben Transport and Metabolism in the Biomimetic Artificial Membrane Permeability Assay (BAMPA) and 3-Day and 21-Day Caco-2 Cell Systems

2006 ◽  
Vol 12 (1) ◽  
pp. 84-91 ◽  
Author(s):  
Mark Lakeram ◽  
David J. Lockley ◽  
David J. Sanders ◽  
Ruth Pendlington ◽  
Ben Forbes
Keyword(s):  
Membrane Permeability ◽  
Systemic Exposure ◽  
In Vitro Models ◽  
Log P ◽  
Hydroxybenzoic Acid ◽  
Artificial Membrane ◽  
Parabolic Relationship ◽  
Permeability Assay

Noncellular and cellular in vitro models for predicting intestinal absorption were used to investigate the transport and metabolism of parabens. The biomimetic artificial membrane permeability assay (BAMPA) membrane was constructed by impregnating a lipid solution on a hydrophobic filter. Caco-2 cells at passage numbers 65 to 80 were cultured in either the accelerated 3-day Biocoat™ system or the standard 21-day Transwell™ cell culture system. Paraben transport across the BAMPA system showed a parabolic relationship. The lowest log P (p-hydroxybenzoic acid) and highest log P compounds (heptyl and octyl parabens) had apparent permeabilities (Papp) less than 1.0 × 10-6 cm/s and Papp was maximal at approximately 8.5 × 10-6cm/s for the intermediate log P (ethylparaben) compound. With the Biocoat™, a similar parabolic relationship was found. In the 21-day Caco-2 cells, the parabens were metabolized by esterases at to p-hydroxybenzoic acid. In conclusion, the in vitro models added complementary insight into the absorption process, such as the transport route, intrinsic permeability, and extent of metabolism of the parabens. This study indicated that presystemic metabolism of orally ingested parabens to the p-hydroxybenzoic acid in the intestine may limit systemic exposure to alkyl-paraben esters in vivo.

scholarly journals Predicting A Drug'S Membrane Permeability: Evolution of a Computational Model Validated with in Vitro Permeability Assay Data

2016 ◽  
Vol 110 (3) ◽  
pp. 330a ◽  
Author(s):  
Timothy S. Carpenter ◽  
M. Windy McNerney ◽  
Nicholas A. Be ◽  
Victoria Lao ◽  
Emma M. Carlson ◽  
...  
Keyword(s):  
Computational Model ◽  
Membrane Permeability ◽  
Permeability Evolution ◽  
Permeability Assay

High throughput artificial membrane permeability assay for blood–brain barrier

2003 ◽  
Vol 38 (3) ◽  
pp. 223-232 ◽  
Author(s):  
Li Di ◽  
Edward H Kerns ◽  
Kristi Fan ◽  
Oliver J McConnell ◽  
Guy T Carter
Keyword(s):  
Membrane Permeability ◽  
Blood Brain Barrier ◽  
High Throughput ◽  
Brain Barrier ◽  
Artificial Membrane ◽  
Blood Brain ◽  
Permeability Assay
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